Salt-precipitation method does not isolate to homogeneity Tamm-Horsfall glycoprotein from urine of proteinuric patients and pregnant women

Objective: Assessment of the degree of purification of Tamm-Horsfall glycoprotein from anomalous urine. Design and methods: Two methods have been compared: the method of Tamm & Horsfall (T&H method) consisting in the precipitation of THP by the addition to urine of NaCl up to 0.58 mol/L and the filtration of urine through a diatomaceous earth filter (DEF method) in which THP is selectively trapped because of its gelation/aggregation tendency. The purity of THP preparations has been evaluated by SDS-PAGE analysis and Western blotting developed with anti immunoglobulin G (IgG) antibodies and antichorionic gonadotropin antibodies. Results: All THPs isolated by T&H method from proteinuric patients were contaminated by IgG and one of the five preparations from pregnant women even by chorionic gonadotropin. A smaller or no contamination was found in THPs isolated by DEF method. Conclusions: Although albumin is the most abundant protein in the anomalous urine, it never appears in THP preparations. The consistent contamination with IgG of THP prepared by salt precipitation-method might be related to the formation of a stable complex between the two proteins.